OsLESV and OsESV1 promote transitory and storage starch biosynthesis to determine rice grain quality and yield.

Transitory starch is an important carbon source in leaves, and its biosynthesis and metabolism are closely related to grain yield and quality. The molecular mechanisms controlling leaf transitory starch biosynthesis and degradation and their effects on rice (Oryza sativa) yield and quality are unclear. Here, we showed that OsLESV and OsESV1, the rice orthologs of AtLESV and AtESV1, are associated with transitory starch biosynthesis in rice. The total starch and amylose contents in leaves and endosperm were significantly reduced, and the final grain quality and yield were compromised in oslesv and osesv1 single and oslesv esv1 double mutants. Further, we found that OsLESV and OsESV1 bind to starch and that this binding depends on a highly conserved C-terminal tryptophan-rich region that acts as a starch-binding domain. Importantly, OsLESV and OsESV1 also interact with the key enzymes of starch biosynthesis, GBSSI, GBSSII, and PPDKB, and maintain their protein stability and activity levels. OsLESV and OsESV1 also assist in targeting GBSSI and GBSSII from plastid stroma to starch granules. Overexpressing GBSSI, GBSSII, and PPDKB partly rescued the phenotypic defects of the oslesv and osesv1 mutants. Thus, we demonstrated that OsLESV and OsESV1 play a key role in regulating the biosynthesis of both leaf transitory starch and endosperm storage starch in rice. These findings enhance our understanding of the molecular mechanisms underlying transitory starch biosynthesis in rice leaves and reveal how transitory starch metabolism affects rice grain quality and yield, providing useful information for the genetic improvement of rice grain quality and yield.

Single-cell RNA sequencing unveils tumor heterogeneity and immune microenvironment between subungual and plantar melanoma.

Acral melanoma (AM) is a subtype of melanoma with high prevalence in East Asians. AM is characterized by greater aggressiveness and lower survival rates. However, there are still fewer studies on immune mechanisms of AM especially subungual melanoma (SM) versus non-subungual melanoma (NSM). In order to explore tumor heterogeneity and immune microenvironment in different subtypes of AM, we applied single-cell RNA sequencing to 24,789 single cells isolated from the SM and plantar melanoma (PM) patients. Aspects of tumor heterogeneity, melanocytes from PM and SM had significant differences in gene expression, CNV and pathways in which tumor-associated such as NF-kb and Wnt were involved. Regarding the immune microenvironment, PM contained more fibroblasts and T/NK cells. The EPHA3-EFNA1 axis was expressed only in cancer-associated fibroblast (CAF) and melanocytes of PM, and the TIGIT-NECTIN2 axis was expressed in both AM subtypes of T/NK cells and melanocytes. Altogether, our study helps to elucidate the tumor heterogeneity in AM subpopulations and provides potential therapeutic targets for clinical research.

Boosting the reversibility of Zn anodes via synergistic cation-anion interface adsorption with addition of multifunctional potassium polyacrylate.

Rechargeable aqueous Zn batteries have the edge in resource reserve, cost, energy and conversion efficiency due to the inherent features of metal Zn anodes. However, the application of Zn-based batteries is being seriously hindered by Zn dendrites and water-induced side-reactions. Here, potassium polyacrylate (K-PAM) is proposed as the electrolyte additive to form a synergistic cation-anion interface on Zn surface. The carboxyl anions and K+ cations are preferentially adsorbed on the Zn surface due to the intrinsic surfactant characteristics, which could homogenize Zn plating and suppress parasitic reactions. The synergistic regulation of K-PAM additive endows the ZnZn symmetric cells with excellent cyclic durability of 1250 h at 1 mA cm-2, which is significantly better than the polyacrylic acid additive only with carboxyl anions. Moreover, trace K-PAM addition into traditional ZnSO4 electrolyte endows the ZnCu batteries with a considerable average Coulombic efficiency of 99.2 %. Additionally, higher capacity retention and excellent cycling stability of ZnVO2 cells further mark K-PAM as a potentially impressive aqueous electrolyte additive for high-performance Zn-based batteries. This work will provide a promising method for the synergistic regulation with cations and anions of electrolyte additives to improve the stability and reversibility of Zn anodes.

Hierarchical Porous Fibers for Intrinsically Thermally Insulated and Self-Sensing Integrated Smart Textile.

Here, stretchable hierarchical porous polyurethane fibers were designed, fabricated, and employed as a three-dimensional hierarchical interconnected framework for conductive networks interwoven with silver nanoparticles and carbon nanotubes. The fiber possessed favorable thermal insulation, strain sensing, and electric heating properties. The core-shell layered porous structure of fiber made the fiber have high heat insulation performance (the difference value of temperature |ΔT| = 3.54, 8.9, and 12.7 °C at heating stage temperatures of 35, 50, and 65 °C) and ultrahigh elongation at break (813%). Importantly, after conductive filler decoration, the fiber could exhibit real-time strain-sensing capacities with a high gauge factor. In addition, the fibers could be heated at low voltage, like an electrical heater. The development of flexible, stretchable, and multifunctional porous fibers had great potential applications in intelligent wearable devices for integrated thermal management, strain sensing, and intrinsic self-warming capability.

Differentiation of parotid pleomorphic adenoma from Warthin tumor using signal intensity ratios on fat-suppressed T2-weighted magnetic resonance imaging.

OBJECTIVE: To investigate the value of magnetic resonance imaging (MRI) signal intensity ratios (SIRs) based on fat-suppressed T2-weighted imaging (FS-T2WI), together with demographic features, MRI anatomical characteristics, and SIRs of histopathological patterns of the tumors, in the differentiation of parotid pleomorphic adenoma (PA) from Warthin tumor (WT). STUDY DESIGN: In total, 90 patients with PA and 56 patients with WT were enrolled in the study. SIRs of tumor to normal parotid gland (SIR-T/P), spinal cord (SIR-T/S), and muscle (SIR-T/M) were calculated. Demographic and radiological features of the 2-patient groups were compared with univariate analysis and multivariate logistic regression analysis. The area under the receiver operating characteristic curve (AUC), sensitivity, and specificity were analyzed to evaluate the utility of SIRs in distinguishing between PA and WT. RESULTS: SIR-T/P exhibited outstanding discriminating ability (AUC = 0.934), SIR-T/S had excellent discrimination (AUC = 0.839), and SIR-T/M showed acceptable discrimination (AUC = 0.728). When SIR-T/P of 1.96 was selected as the cutoff value, sensitivity and specificity were 0.756 and 0.982, respectively. SIR-T/P, age, sex, and number of lesions were identified as independent predictors by multivariate logistic regression analysis. Differences in SIRs between histopathological patterns were significant. CONCLUSION: SIR-T/P based on FS-T2WI is an effective discriminator in the differential diagnosis between PA and WT. Age, sex, and number of lesions provided additional value in differentiation.

Wild imitating vs greenhouse cultivated Dendrobium huoshanense: Chemical quality differences.

Dendrobium huoshanense (D. huoshanense) has been used as functional food supplements and herbal medicines for preventing and managing diseases with a long history in China. Due to its endangered natural resources and huge demand, people tend to cultivate D. huoshanense to protect this species. However, the quality of wild and cultivated herbs of the same species may change. This work quantified and compared the main quality traits and chemical components of wild imitating and greenhouse cultivated D. huoshanense with different growth years. As a result, wild and cultivated D. huoshanense had similar chemical composition, but there are significant differences in the content of many ingredients (polysaccharides, flavonoids, nucleosides, bibenzyls, lignans and volatile compounds). And the contents of many of these components increased with growing years. In addition, multivariate statistical analyses have been applied to classify and evaluate samples from different cultivation modes according to these components. In conclusion, our results demonstrated that the overall quality of greenhouse cultivated D. huoshanense was not as good as wild-grown, but this mode can be a promising and sustainable way of producing D. huoshanense.

Hepatic steatosis using ultrasound-derived fat fraction: First technical and clinical evaluation.

OBJECTIVES: To explore the technical and clinical evaluation of ultrasound-derived fat fraction (UDFF) measurement in adult patients in whom fatty liver was suspected. MATERIALS AND METHODS: In this prospective study, 41 participants were initially enrolled in our hospital between October 2022 and December 2022 and received UDFF assessment using Siemens ACUSON Sequoia system equipped with DAX transducer. UDFF measurement was performed three times to obtain UDFF values from each imaging location (V hepatic segment and VIII hepatic segment) per participant, and the depth (skin-to-capsule distance) was automatically measured. The echogenicity of liver tissue in B mode ultrasound (BMUS) was compared to the normal kidney tissue, and fatty liver was graded as mild (Grade 1), moderate (Grade 2), and severe (Grade 3). The median of the acquired overall median UDFF values was used for statistical analysis. All ultrasound examinations were performed by one of two radiologists (with 20 and 10 years of liver ultrasound imaging experience). RESULTS: Finally, UDFF measurement was successfully performed on 38 participants to obtain valid values, including 21 men with a median age of 40.0 years (interquartile range [IQR]: 23.0 - 58.5) and 17 women with a median age of 60.0 years (IQR: 29.5 - 67.0). Fatty liver was diagnosed by BMUS features in 47.4% (18/38) participants. Among all participants, the median UDFF value was 7.0% (IQR: 4.0 - 15.6). A significant difference in UDFF values was found between participants with fatty liver and without fatty liver (U = 7.0, P < 0.001), and UDFF values elevated as the grade of the fatty liver increased (P < 0.001). The median UDFF values from the three UDFF measurements obtained during each ultrasound examination showed excellent agreement (ICC = 0.882 [95% confidence interval: 0.833 - 0.919]). The Spearman correlation of UDFF values in different depths was moderate, with a rs value of 0.546 (P < 0.001). No significant differences in UDFF values were found between V hepatic segment and VIII hepatic segment (U = 684.5, P = 0.697). CONCLUSIONS: UDFF provides a novel non-invasive imaging tool for hepatic steatosis assessment with excellent feasibility.

Metal-Organic Framework Glass as a Functional Filler Enables Enhanced Performance of Solid-State Polymer Electrolytes for Lithium Metal Batteries.

Polymers are promising candidates as solid-state electrolytes due to their performance and processability, but fillers play a critical role in adjusting the polymer network structure and electrochemical, thermal, and mechanical properties. Most fillers studied so far are anisotropic, limiting the possibility of homogeneous ion transport. Here, applying metal-organic framework (MOF) glass as an isotropic functional filler, solid-state polyethylene oxide (PEO) electrolytes are prepared. Calorimetric and diffusion kinetics tests show that the MOF glass addition reduces the glass transition temperature of the polymer phase, improving the mobility of the polymer chains, and thereby facilitating lithium (Li) ion transport. By also incorporating the lithium salt and ionic liquid (IL), Li-Li symmetric cell tests of the PEO-lithium salt-MOF glass-IL electrolyte reveal low overpotential, indicating low interfacial impedance. Simulations show that the isotropic structure of the MOF glass facilitates the wettability of the IL by enhancing interfacial interactions, leading to a less confined IL structure that promotes Li-ion mobility. Finally, the obtained electrolyte is used to construct Li-lithium iron phosphate full batteries that feature high cycle stability and rate capability. This work therefore demonstrates how an isotropic functional filler can be used to enhance the electrochemical performance of solid-state polymer electrolytes.

Altered fatty acid composition confers improved drought acclimation in maize.

Drought induces alteration in membrane lipid composition in plants; however, still little is known about whether membrane lipid remodeling plays a role in plant drought acclimation, including both drought tolerance and recovery, especially in crops. Here, we imposed natural progressive drought and re-watering in 18 maize genotypes at the seedling stage, and analyzed the physiological responses, drought tolerance and drought acclimation capabilities, contents of lipids, and fatty acid compositions. The results showed that drought caused significant reductions in shoot dry weight, relative water content, Fv/Fm, total lipid content, and double bond index (DBI) in most genotypes, while re-watering partially recovered these reductions. Meanwhile, the total lipid content, fatty acid composition, and DBI were also changed obviously in response to drought and re-watering. In order to explore the relationship between membrane lipid change and plant drought response, we did a principal component analysis. The results showed that C18:3 fatty acid contributed greatly to drought tolerance, and C16:2 and C16:3 fatty acids were more responsible for drought recovery. Meanwhile, DBI showed significant positive correlations with shoot dry weight and relative water content, but a negative association with lipid peroxidation, and more importantly, DBI was important for both drought tolerance and recovery. These alterations in membrane lipid composition may facilitate increasing membrane fluidity and decreasing membrane damage, thus maintaining the high photosynthetic capability under drought. Our results suggest that lipid remodeling is important for drought tolerance and recovery in crops, and different fatty acid species have different roles in crop drought acclimation.

Characteristics of common pathogens of urogenital tract among outpatients in Shanghai, China from 2016 to 2021.

Background: Ureaplasma urealyticum, Chlamydia trachomatis, and Neisseria gonorrhoeae are the prevalent causes of several genital diseases worldwide; however, their characteristics in different genders have not been well documented in Shanghai. The aim of this study is to describe the prevalence of common pathogens among outpatients, considering variations by gender and age. Methods: From January 1, 2016, to December 31, 2021, the urogenital swabs of 16216 outpatients aged 3-95 years from two general hospitals in Shanghai were collected. All participants' swabs were investigated for U. urealyticum, C. trachomatis, and N. gonorrhoeae by isothermal RNA-based simultaneous amplification and testing. The basic information of all participants was also recorded, including age and gender. The chi-square test was used to compare the prevalence between different genders, age groups, and infection patterns. Results: There were 5,744 patients (35.42%) with positive samples whose ages ranged from 7 to 80 years (33.23 ± 8.63 years), and 62.14% of them were women. The most common pathogen detected was U. urealyticum (85.08%). The highest prevalence rate of all three pathogens was found in patients aged ≤ 20 years (40.53%, 95% confidence intervals [CI]: 33.80%-47.63%). The prevalent rate of U. urealyticum was higher in men (33.36%, 95% CI: 32.19%-34.55%). The overall prevalence rates of U. urealyticum, C. trachomatis, and N. gonorrhoeae were 30.14% (95% CI: 29.44%-30.85%), 6.00% (95% CI: 5.64%-6.38%), and 2.10% (95% CI: 1.89%-2.33%). Conclusions: Ureaplasma urealyticum was the most prevalent pathogen in the population, and its prevalence decreased with age. Young men aged ≤ 20 years were more frequently infected. Regular screening for sexually transmitted pathogens in different genders and age groups are warranted, particularly in young men.

A glucose-mediated antibiotic resistance metabolic flux from glycolysis, the pyruvate cycle, and glutamate metabolism to purine metabolism.

Introduction: Bacterial metabolic environment influences antibiotic killing efficacy. Thus, a full understanding for the metabolic resistance mechanisms is especially important to combat antibiotic-resistant bacteria. Methods: Isobaric tags for relative and absolute quantification-based proteomics approach was employed to compare proteomes between ceftazidime-resistant and -sensitive Edwarsiella tarda LTB4 (LTB4-RCAZ and LTB4-S, respectively). Results: This analysis suggested the possibility that the ceftazidime resistance mediated by depressed glucose is implemented through an inefficient metabolic flux from glycolysis, the pyruvate cycle, glutamate metabolism to purine metabolism. The inefficient flux was demonstrated by the reduced expression of genes and the decreased activity of enzymes in the four metabolic pathways. However, supplement upstream glucose and downstream guanosine separately restored ceftazidime killing, which not only supports the conclusion that the inefficient metabolic flux is responsible for the resistance, but also provides an effective approach to reverse the resistance. In addition, the present study showed that ceftazidime is bound to pts promoter in E. tarda. Discussion: Our study highlights the way in fully understanding metabolic resistance mechanisms and establishing metabolites-based metabolic reprogramming to combat antibiotic resistance.

Confinement Effect and 3D Design Endow Unsaturated Single Ni Atoms with Ultrahigh Stability and Selectivity toward CO2 Electroreduction.

Developing single-atomic catalysts with superior selectivity and outstanding stability for CO2 electroreduction is desperately required but still challenging. Herein, confinement strategy and three-dimensional (3D) nanoporous structure design strategy are combined to construct unsaturated single Ni sites (Ni-N3 ) stabilized by pyridinic N-rich interconnected carbon nanosheets. The confinement agent chitosan and its strong interaction with g-C3 N4 nanosheet are effective for dispersing Ni and restraining their agglomeration during pyrolysis, resulting in ultrastable Ni single-atom catalyst. Due to the confinement effect and structure advantage, such designed catalyst exhibits a nearly 100% selectivity and remarkable stability for CO2 electroreduction to CO, exceeding most reported state-of-the-art catalysts. Specifically, the CO Faradaic efficiency (FECO ) maintains above 90% over a broad potential range (-0.55 to -0.95 V vs. RHE) and reaches a maximum value of 99.6% at a relatively low potential of -0.67 V. More importantly, the FECO is kept above 95% within a long-term 100 h electrolyzing. Density functional theory (DFT) calculations explain the high selectivity for CO generation is due to the high energy barrier required for hydrogen evolution on the unsaturated Ni-N3 . This work provides a new designing strategy for the construction of ultrastable and highly selective single-atom catalysts for efficient CO2 conversion.

Geospatial analysis of the hospitalisation rate of patients with rheumatoid arthritis in Hunan: a cross-sectional Chinese study.

OBJECTIVE: Little is known about spatial variability of hospitalisation rate (HR) of patients with rheumatoid arthritis (RA) worldwide, especially in China. METHODS: A cross-sectional study was conducted among patients with RA admitted to hospitals in Hunan Province. Global Moran's I and local indicators of spatial association were used to explore the geospatial pattern of the HR of patients with RA. Generalised estimating equation analysis and geographically weighted regression were used to identify the potential influencing factors of the HR of patients with RA. RESULTS: There were a total of 11 599 admissions, and the average HR was 1.57 per 10 000 population in Hunan. We detected different cluster patterns of the HR among patients with RA by local indicators of spatial association. Age, ethnicity, average temperature, average temperature range, average rainfall, regions, gross domestic product per capita, and doctors and hospitals per 10 000 people were risk factors for the HR. However, only average temperature, gross domestic product per capita and hospitals per 10 000 people showed different regression coefficients on the HR in different counties. The increase in hospitals increased the probability of HR from east to west in Hunan with a positive coefficient, while temperature decreases increased the risk of HR from south to north negatively. Similarly, the growth of gross domestic product per capita decreased the probability of HR from southwest to northeast. CONCLUSION: A non-random spatial distribution of the HR of patients with RA was demonstrated in Hunan, and average temperature, gross domestic product per capita and hospitals per 10 000 people showed different regression coefficients on the HR in different counties. Our study indicated that spatial and geostatistics may be useful approaches for further study among patients with RA.

Molecular identification and functional analysis of chitinase genes reveal their importance in the metamorphosis of Sarcophaga peregrina (Diptera: Sarcophagidae).

Chitinases play a crucial role in insect metamorphosis by facilitating chitin degradation. Sarcophaga peregrina (Robineau-Desvoidy, 1830) (Diptera: Sarcophagidae) is a typical holometabolous insect and an important hygiene pest that causes myiasis in humans and other mammals and acts as a vector for various parasitic agents, including bacteria, viruses, and parasites. Enhancing the understanding of the metamorphosis in this species has significance for vector control. In this study, we identified a total of 12 chitinase genes in S. peregrina using bioinformatic analysis methods. Based on transcriptome data, SpIDGF2 and SpCht10 were selected for further functional investigation. The down-regulation of these genes by RNA interference led to developmental delays, disruptions in molting, and differences in cuticle composition during the pupal stage. These findings underscore the pivotal role of chitinase genes in the metamorphic process and offer valuable insights for effective control strategies.

Identification and Analysis of a Four-Gene Set for Diagnosing SFTS Virus Infection Based on Machine Learning Methods and Its Association with Immune Cell Infiltration.

Severe Fever with thrombocytopenia syndrome (SFTS) is a highly fatal viral infectious disease that poses a significant threat to public health. Currently, the phase and pathogenesis of SFTS are not well understood, and there are no specific vaccines or effective treatment available. Therefore, it is crucial to identify biomarkers for diagnosing acute SFTS, which has a high mortality rate. In this study, we conducted differentially expressed genes (DEGs) analysis and WGCNA module analysis on the GSE144358 dataset, comparing the acute phase of SFTSV-infected patients with healthy individuals. Through the LASSO-Cox and random forest algorithms, a total of 2128 genes were analyzed, leading to the identification of four genes: ADIPOR1, CENPO, E2F2, and H2AC17. The GSEA analysis of these four genes demonstrated a significant correlation with immune cell function and cell cycle, aligning with the functional enrichment findings of DEGs. Furthermore, we also utilized CIBERSORT to analyze the immune cell infiltration and its correlation with characteristic genes. The results indicate that the combination of ADIPOR1, CENPO, E2F2, and H2AC17 genes has the potential as characteristic genes for diagnosing and studying the acute phase of SFTS virus (SFTSV) infection.

A rapid and high-throughput multiplex genetic detection assay for detection, semi-quantification and virulence genotyping of Helicobacter pylori in non-invasive oral samples.

Aim: This study established a high-throughput multiplex genetic detection assay (HMGA) for rapid identification, semi-quantification and virulence analysis of Helicobacter pylori directly from the clinical non-invasive oral samples. Methods: The gastric mucosa and oral samples were collected from 242 patients in Shanghai from 2021 to 2022. All the samples were detected by routine clinical tests for H. pylori and Sanger sequenced for inconsistent results. A new multiplex PCR assay providing results within 4 hours was designed and optimized involving fluorescent dye-labeled specific primers targeted 16S rRNA gene, semi-quantitative gene ureC and 10 virulence genes of H. pylori. Semi-quantification was carried out by simulating the serial 10-fold dilutions of positive oral samples, and the H. pylori loads in different clinical samples were further compared. The mixed plasmids of virulence genes vacA s1, vacA m1 and vacA m2 were used to evaluate the performance on different genotypes. The consistency of 10 virulence genes in gastric mucosa, saliva, mouthwash and dental plaque of H. pylori-positive patients was compared. Results: The non-invasive HMGA was highly specific for detection of all 12 targets of H. pylori and human internal reference gene ß-globin, and the sensitivity to all target genes could reach 10 copies/µL. Compared with routine clinical tests and sequencing, non-invasive HMGA has a high level (>0.98) of sensitivity, specificity, accuracy, PPV, NPV and kappa coefficient for direct detection of H. pylori in oral samples. Moreover, by detecting peak area levels of ureC, it was confirmed that the H. pylori loads in gastric mucosa were significantly higher than those of the three kinds of oral samples (p<0.05). We also found that 45.0% (91/202) of patients had different H. pylori virulence genes in different oral samples. The concordance of positive detection rates of each virulence gene between saliva and gastric mucosa was more than 78% (p<0.05). Conclusion: The non-invasive HMGA proved to be a reliable method for the rapid H. pylori identification, semi-quantification and detection of 10 virulence genes directly in oral samples, providing a new idea for non-invasive detection of H. pylori.

Identification of Intestinal Microbial Community in Gallstone Patients with Metagenomic Next-Generation Sequencing.

Gallstone disease (GD) is one of the most common gastrointestinal diseases worldwide. Nowadays, intestinal microbiota are thought to play important roles in the formation of gallstones. In our study, human fecal samples were extracted for metagenomic next-generation sequencing (mNGS) on the Illumina HiSeq platform, followed by bioinformatics analyses. Our results showed that there was a particular intestinal micro-ecosystem in GD patients. In contrast to healthy people, the sequences of Bacteroidetes, Bacteroides and Thetaiotaomicron were obviously more abundant in GD patients at phylum, genus and species levels, respectively. On the other hand, the glycan metabolism and drug resistance, especially for the ß-lactams, were the most profound functions of gut microbes in GD patients compared to those in normal subjects. Furthermore, a correlation analysis drew out that there existed a significant relationship between the serum levels of biochemical indicators and abundances of intestinal microbes in GD patients. Our results illuminate both the composition and functions of intestinal microbiota in GD patients. All in all, our study can broaden the insight into the potential mechanism of how gut microbes affect the progression of gallstones to some extent, which may provide potential targets for the prevention, diagnosis or treatment of GD.

Knockout of CLTC gene reduces but not completely block SFTSV infection.

Clathrin is a key protein for viruses to enter host cells. Previous studies often use clathrin inhibitors or gene knockdown technology to partially inhibit the function of clathrin, but whether SFTSV can infect host cells without clathrin expression remains unclear. In this research, a clathrin heavy chains (CLTC) knockout A549 cell line was established by CRISPR/Cas9 technology, and the knockout of CLTC was verified by PCR, Western blot, immunofluorescence and T7E1 analysis. The off-target effect was evaluated by PCR combined with Sanger sequencing. Furthermore, this research verified that SFTSV infection was significantly inhibited, but not completely blocked, due to the deletion of CLTC protein. Our research also found that lipid raft inhibitor Filipin, other than macropinocytosis inhibitor EIPA, could significantly reduce SFTSV infection, and the inhibition was more obviously observed when Filipin was used in CLTC knockout cells. These result indicated that clathrin-dependent and lipid raft mediated endocytosis are the major two mode used by SFTSV entry. In conclusion, this study constructed a CLTC knockout cell line, which, for the first time, established a cell model for the study of the function of CLTC protein, and provided direct evidence that SFTSV pendent could still infect cells without clathrin. Additionally, we confirmed that lipid raft mediated endocytosis, as a clathrin-independent pathway, could be another key mode for SFTSV entry.